In vitro cytotoxicity evaluation of endodontic irrigants combined with cationic peptide.

Abstract

Aim: The aim of this study was to assess the cytotoxicity of endodontic irrigants 2% chlorhexidine (CHX) and ethylenediaminetetraacetic acid (EDTA) combined with cationic peptide DJK-5 on L929 mouse subcutaneous fibroblast cells.

Subjects and methods: L929 cells were cultured and treated with 2% CHX, 2% CHX + DJK-5, 8.5% EDTA, 8.5% EDTA + DJK-5, and DJK-5 alone (DJK-5 peptide concentration-10 μg/mL) for 24 and 48 h. The cytotoxicity of the endodontic irrigants was determined using methyl thiazolyl tetrazolium assay and Live/Dead assay using acridine orange and ethidium bromide dual fluorescence staining.

Statistical analysis used: The Shapiro-Wilk test was used to verify the normality of the data within each group at two different time points. Parametric tests were used for the comparison. Paired sample t-test was used for the comparison of the cytotoxicity of irrigants over time. One-way ANOVA was employed for the comparison of the cytotoxicity of irrigants. Tukey's test was used for pairwise comparison between groups. SPSS software version 27 was used for the data analyses.

Results: CHX exhibited slight cytotoxicity at 24 h and higher cytotoxicity at 48 h (P < 0.05). EDTA showed better cell viability than CHX at both time points. In combination groups, the EDTA + DJK-5 group showed lower cytotoxicity at both time points (P > 0.05). The combination of CHX with DJK-5 reduced the cytotoxicity of CHX (P < 0.05).

Conclusions: The cytotoxicity of endodontic irrigants was found to be time dependent. CHX was found to be highly cytotoxic at 2% concentration. DJK-5 was found to have the least cytotoxicity, and it significantly reduced the cytotoxicity of CHX and EDTA.